Human primary colon cancer cell separation and culture method

A technology of human colon cancer cells and colon cancer cells, which is applied in the field of cell culture of modern biotechnology, can solve the problems of unfavorable practical application, low cell purity, long culture time, etc., and achieve good cell growth status, high cell purity, Make simple effects

Inactive Publication Date: 2018-05-25
JIANGYIN CHI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, most of the established methods for the isolation and culture of human colon cancer cells are based on tissue block culture. The isolated cells are not high in purity, small in number, and have a

Method used

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  • Human primary colon cancer cell separation and culture method
  • Human primary colon cancer cell separation and culture method
  • Human primary colon cancer cell separation and culture method

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Embodiment Construction

[0035] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0036] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0037] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0038] The experimental instruments and reagents used in this experiment are as follows:

[0039] A set of surgical instruments, CCK-8 cell counting box (Sigma, USA), enzyme-linked immunosorbent assay kit (R&D, USA), inverted microscope (XDS-1A, Shanghai), fluorescence microscope (Leica, USA), cryogenic centrifugation Machine (TD24B-WS, Shanghai), ultra-low temperature refrigerator (Zhongke Meiling), pipette gun (Eppendorf, USA), electronic analytical balance (Sartorius, U...

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Abstract

The invention provides a human primary colon cancer cell separation and culture method. The method comprises the following steps of (1) cutting a fresh colon cancer tissue under an aseptic condition,sealing at 4 DEG C and then carrying back to a lab; (2) disinfecting a tissue processing device; (3) using a pair of operating scissors for separating the colon cancer tissue, placing the tissue intoa precooled sterile PBS liquid containing double antibodies, and washing multiple times to remove blood vessels, fat and necrotic tissues; (4) cutting up the tissues, and using preheated pancreatin and IV type collagenase for digesting respectively; (5) terminating digestion, centrifuging, and removing a supernatant; (6) using trypan-blue for dyeing a cell filter liquor to detect cell activity; (7) after carrying out cell counting, inoculating into a coated culture flask; (8) culturing at 37 DEG C in an environment with 5 percent of CO2; (9) purifying the cells; (10) subculturing the tissues;(11) observing cell morphology; (12) determining cell survival rate; (13) carrying out immunological identification. The invention provides the human primary colon cancer cell separation and culture method which is simple to operate, the number of the obtained cells is large, the survival rate is high, the method is the ideal human primary colon cancer cell primary separation and culture method, and a reliable cell resource is provided for experiments.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, and specifically relates to a method for separating and culturing primary human colon cancer cells. Background technique [0002] Colon cancer is one of the most common clinical malignant tumors and lethal factors. Although new chemotherapy drugs have emerged in recent years, their complete cure rate and long-term survival rate are still very low. This situation may be related to tumor stem cells. Small amount of CD133 found in colon cancer + Cells, which are able to maintain colon cancer, are closely related to drug resistance and treatment of colon cancer. Therefore, it is of great significance to find such stem cells of colon cancer for the treatment of colon cancer. [0003] At present, cell lines are mostly used in the research of human colon cancer, but long-term subculture in vitro will change the cell characteristics and protein expression, which may interfe...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2501/33C12N2509/00C12N2533/32
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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